This antibody was isolated by affinity chromatography using antigen coupled to agarose beads.

克隆

多克隆

同种型

IgG

常规说明

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to use products containing Alexa Fluor® dyes for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com

The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab7291, 1µg/ml) and (ab16048, 1µg/ml) overnight at +4°C. The secondary antibodies were ab150115 Alexa Fluor® 647 (red) goat anti-mouse IgG (H+L) used at 2µg/ml for 1h and ab150077 Alexa Fluor® 488 (green) goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei.

Overlay histogram showing Jurkat cells stained with ab16669 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Overlay histogram showing HeLa cells stained with ab32356 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32356, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Primary antibody anti-Glycine (ab9442) was used at 1 in 500. Secondary antibody Goat anti-Rabbit (ab150077) was used at 1 in 1000, for 2 hours at room temperature. Following 3 washes there was still strong staining and little background noise.

Following fixation in 4% PFA, the cells were assessed for Sall4 expression using 1:100 dilution of the primary antibody (ab29112) in 1% serum, 0.1% triton, 0.1% BSA in PBS, followed by detection using goat polyclonal rabbit IgG Alexa 488 (ab150077) at 1:200. The results show that nuclear Sall4 (green) was clearly observed. An isotype control IgG was run in parallel and showed no positive staining (not shown here).

Cells were fixed in 4% PFA, permeabilized using 0.1% Triton X-100, blocked with 1% Goat serum, 0.1% BSA in PBS for 30 minutes at RT then incubated with ab124297 at a 1/100 dilution or Rabbit IgG Isotype control for 2 hours at RT. The secondary used was a Goat polyclonal Secondary Antibody to Rabbit IgG – H&L Alexa Fluor 488 (ab150077) used at 1/200 dilution.